Primary Cell Culture of Live Neurosurgically Resected Aged Adult Human Brain Cells and Single Cell Transcriptomics

Publication date: 17 January 2017
Source:Cell Reports, Volume 18, Issue 3
Author(s): Jennifer M. Spaethling, Young-Ji Na, Jaehee Lee, Alexandra V. Ulyanova, Gordon H. Baltuch, Thomas J. Bell, Steven Brem, H. Isaac Chen, Hannah Dueck, Stephen A. Fisher, Marcela P. Garcia, Mugdha Khaladkar, David K. Kung, Timothy H. Lucas, Donald M. O’Rourke, Derek Stefanik, Jinhui Wang, John A. Wolf, Tamas Bartfai, M. Sean Grady, Jai-Yoon Sul, Junhyong Kim, James H. Eberwine
Investigation of human CNS disease and drug effects has been hampered by the lack of a system that enables single-cell analysis of live adult patient brain cells. We developed a culturing system, based on a papain-aided procedure, for resected adult human brain tissue removed during neurosurgery. We performed single-cell transcriptomics on over 300 cells, permitting identification of oligodendrocytes, microglia, neurons, endothelial cells, and astrocytes after 3 weeks in culture. Using deep sequencing, we detected over 12,000 expressed genes, including hundreds of cell-type-enriched mRNAs, lncRNAs and pri-miRNAs. We describe cell-type- and patient-specific transcriptional hierarchies. Single-cell transcriptomics on cultured live adult patient derived cells is a prime example of the promise of personalized precision medicine. Because these cells derive from subjects ranging in age into their sixties, this system permits human aging studies previously possible only in rodent systems.

Graphical abstract

Teaser

Human CNS disease is best studied in human neuronal cells. Singh et al. have succeeded in performing long-term primary cell culture and single-cell transcriptomics of human adult brain cells, including neurons from patients up to 67 years of age.


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